Abstract
Accurate, sensitive, and specific detection of molecular markers in intraocular fluid will facilitate the early discovery, diagnosis, and intervention of eye diseases. In this study, a total of 168 participants were recruited and divided into two distinct cohorts: discovery and verification. In the discovery phase, proteomic analysis identified MCP-1 in aqueous humor as a potential molecular marker for blinding eye disease. We further developed a molecular detection technology for the marker based on biolayer interference sensing. The technology utilizes a sandwich strategy with one-to-one pairing of two different biorecognition molecules for MCP-1. It also incorporates automation, high throughput, and real-time monitoring, achieving highly selective recognition and accurate analysis of MCP-1. It demonstrates a low detection limit (0.16 pM), good reliability (R(2) = 0.995), and a wide analytical range (0.244-1000 pM) for MCP-1 in human aqueous humor samples. Crucially, in the verification phase with 150 subjects, the technology achieved a high detection rate (95.0%) for patients with age-related macular degeneration and high myopia cataract in under 30 min, and was able to further differentiate between them with a specificity of 86.0%. Therefore, the developed molecular detection technology may provide a robust, convenient, and valuable solution for widespread screening, early discovery, and differential diagnosis of blinding eye diseases.